PubMed · 5098254
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L E Hammarstrom, S U Toverud, J S Hanker. 1971. Naphthylamidase in ameloblasts during enamel maturation.. https://doi.org/10.1007/bf02062616
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The amelogenin gene is tightly regulated at the temporal and spatial level in accord with the developmental requirement for tooth formation. Previous studies have shown that CCAAT/enhancer-binding protein alpha (C/EBPalpha) is a transactivator of the mouse X-chromosomal amelogenin gene. C/EBPalpha contains four highly conserved regions (CR) named CR1, CR2, CR3, and CR4. Transient transfection assays showed that CR2 in isolation had an exceptional capacity to enhance transcription from the 2.3 kb mouse amelogenin promoter. The remaining conserved regions of C/EBPalpha, either in isolation or in selected combinations, were less effective in amelogenin transactivation than the full length C/EBPalpha. Msx2 has previously been shown to antagonize C/EBPalpha through protein-protein interactions with C/EBPalpha, and the carboxyl-terminus of Msx2 is required for protein-protein interactions. Co-immunoprecipitation analyses identified that the carboxyl-terminal domain (residues 218-359) of C/EBPalpha is required for the C/EBPalpha-Msx2 protein-protein interactions.
Dentin sialophosphpoprotein (Dspp) is transiently expressed in the early stage of secretory ameloblasts. The secretion of ameloblast-derived Dspp is short-lived, correlates to the establishment of the dentinoenamel junction (DEJ), and is consistent with Dspp having a role in producing the specialized first-formed harder enamel adjacent to the DEJ. Crack diffusion by branching and dissipation within this specialized first-formed enamel close to the DEJ prevents catastrophic interfacial damage and tooth failure. Once Dspp is secreted, it is subjected to proteolytic cleavage that results in two distinct proteins referred to as dentin sialoprotein (Dsp) and dentin phosphoprotein (Dpp). The purpose of this study was to investigate the biological and mechanical contribution of Dsp and Dpp to enamel formation. Transgenic mice were engineered to overexpress either Dsp or Dpp in their enamel organs. The mechanical properties (hardness and toughness) of the mature enamel of transgenic mice were compared with genetically matched and age-matched nontransgenic animals. Dsp and Dpp contributions to enamel formation greatly differed. The inclusion of Dsp in bulk enamel significantly and uniformly increased enamel hardness (20%), whereas the inclusion of Dpp weakened the bulk enamel. Thus, Dsp appears to make a unique contribution to the physical properties of the DEJ. Dsp transgenic animals have been engineered with superior enamel mechanical properties.
Sonic hedgehog (Shh), one of the essential molecules for embryogenesis and organogenesis, is strongly expressed in the enamel knot, which represents the signaling center for odontogenesis due to the presence of essential secretory molecules. Here, we investigated the effects of Shh on ameloblast differentiation using a mouse ameloblast-lineage cell line (ALC). In the presence of Shh, the expression of amelogenin and ameloblastin was up-regulated, and their promoter activities were also elevated. Subcutaneous implantation of Shh-treated ALC into the dorsa of C57/Bl6J mouse caused increased expression of amelogenin and ameloblastin. Forced expression of Gli1, a major transcription factor in Shh signaling, up-regulated both amelogenin and ameloblastin, while Gli1-specific RNAi blocked their expression. These results suggest that Shh directly promotes ALC differentiation, and that Gli1 may be responsible for the signal mediation of Shh.